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Principle

This kit uses the Sandwich-ELISA principle. The microtiter plate strips has been pre-coated with an affinity purified antibody to Human TNF-α. Standards or samples containing Human TNF-α are added to the plate and reacted with capture antibody. A second anti-Human TNF-α antibody labelled biotin is then added and binds to Human TNF-α captured on the plate. After that, Streptavidin- Horseradish Peroxidase(SA-HRP) is added to form a sandwich complex of solid phase antibody-Human TNF-α-biotin labeled antibody-SA-HRP. And then, TMB substrate solution is added to all wells and incubated. An enzyme-catalyzed reaction generates a blue color in the solution, thereafter, stop solution is added to stop the substrate reaction and the color turns yellow. The yellow solution is read at a wavelength of 450nm. The concentration of Human TNF-α in the samples is then calculated from the OD value by establishing a standard curve.

Specification

Product name QuanticonTM Human TNF-α(Tumor Necrosis Factor Alpha) ELISA Kit
Cat no ABQ-ES96-TNFAhu
Applications ELISA
Reactivity Human
Standard Range 7.81-500pg/mL
Sensitivity 3.91 pg/mL.
Sample Type Serum, plasma and other biological fluids, Sample
Sample Volume 100 ul
Alternative DIF, TNF-alpha, TNFA, TNFSF2
Assay Type Sandwich
Assay Duration 3.5H
Detection Wavelength 450nm
Storage If the kit is not open Store whole kit: 2-8°C (short time storage valid for 6 months) ; -20°C (long- term storage , valid for 1 year.

Precision

Mean coefficient of variation for Intra-Assay and Inter-Assay: 3 samples with low, middle and high level concentration were tested for repeat multiple times, respectively.

Item Intra-assay Precision Intra-assay Precision
Sample number 3 3
Replicate 9 18
CV(%) 5 8

 

Rate of recovery

The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Matrix type Recovery Range (%) Average (%)
Serum (n=5) 80-90 85
EDTA plasma (n=5) 84-106 100
Cell culture media (n=5) 87-93 90
“Quanticon” Human TNF-α ...

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